title: Inhibiting autophagy by shRNA knockdown: cautions and recommendations.
abstract: A glance through Autophagy or any other journal in this field shows that it is very common to block autophagy by RNA interference-based knockdown of ATG mRNAs in mammalian cell lines. Our lab's experience is that this approach can easily make for failed experiments because good knockdown of even essential autophagy regulators does not necessarily mean you will get good inhibition of autophagy, and, over time, cells can find ways to circumvent the inhibitory effects of the knockdown.
authors.affiliation: Department of Pharmacology; University of Colorado School of Medicine; Anschutz Medical Campus; Aurora, CO USA.
authors.forename: Leah
authors.initials: L
authors.lastname: Staskiewicz
authors.affiliation: Department of Pharmacology; University of Colorado School of Medicine; Anschutz Medical Campus; Aurora, CO USA.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Thorburn
authors.affiliation: Department of Pharmacology; University of Colorado School of Medicine; Anschutz Medical Campus; Aurora, CO USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Morgan
authors.affiliation: Department of Pharmacology; University of Colorado School of Medicine; Anschutz Medical Campus; Aurora, CO USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Thorburn
mesh_terms: D001343:Autophagy / Q000235:genetics*; D055785:Gene Knockdown Techniques / Q000379:methods; D006801:Humans; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference / Q000502:physiology; D034741:RNA, Small Interfering / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering
keywords: ATG5; RNA interference; autophagy; knockdown; shRNA